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YTXPlanned District Craft (US Navy)
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Certified reference materials (CRMs) of OA (13.7 [+ or -] 0.6 [micro]g [ml.sup.-1]), DTX-1 (15.1 [+ or -] 1.1 [micro]g [ml.sup.-1]), DTX-2 (7.8 [+ or -] 0.4 [micro]g [ml.sup.-1]), PTX-2 (4.40 [+ or -] 0.13 [micro]g [ml.sup.-1]), AZA-1 (1.24 [+ or -] 0.07 [micro]g [ml.sup.-1]), AZA-2 (1.28 [+ or -] 0.05 [micro]g [ml.sup.-1]), AZA-3 (1.04 [+ or -] 0.04 [micro]g [ml.sup.-1]), YTX (5.6 [+ or -] 0.2 [micro]g [ml.sup.-1]), and hYTX (5.8 [+ or -] 0.3 [micro]g [ml.sup.-1]) were purchased from the National Research Council, Institute for Marine Biosciences (NRC CNRC, Halifax, Canada).
Table 2 shows the twelve marker genes and three reference genes that were selected from the whole genome array studies where undifferentiated Caco-2 cells were exposed to OA, DTX-1, and AZA-1 (Bodero et al., 2018b) and to YTX and PTX-2 (unpublished data).
For the identification of these toxins, commercial standards of okadaic acid (OA), dinophysistoxin-1 (DTX1), dinophysistoxin-2 (DTX2), pectenotoxin-2 (PTX2), spirolide-1 (SPX1), azaspiracids (AZP1, 2,3) and yessotoxins (YTX) were used.
Mass (m/z) of the standard YTX ions (top) and in the mollusk extracts (lower).
The following negative transitions (precursor ion > product ion) were used for quantifying and confirming (confirming ions shown in parentheses) OA and (DTX-2): m/z 803.5 > 255.0 (803.5 > 113.0); (DTX-1): m/z 817.2 > 255.0 (817.2 > 113.0); YTX: m/z 1,141.5 > 1,061.7 (1,141.5 > 855.5).
No OA, DTX-2, AZA, and YTX were detected in the Dinophysis samples.
B9: (adt) [conjunction] (bdc) [conjunction] (a [not equal to] d) [right arrow] [there exists]x[there exists]y[(abx) [conjunction] (acy) [conjunction] (ytx)].
Standard solutions OA (14.3 [micro]g/mL), PTX2 (8.6 [micro]g/mL), and YTX (5.3 [micro]g/mL) were obtained, in methanol, from the Institute for Marine Biosciences of the National Research Council (Halifax, Nova Scotia, Canada).
For identification of toxins, SRM chromatograms were recorded in negative ionization mode for the transitions m/z 803.5 [right arrow] 255.5 (OA), m/z 817.5 [right arrow] 255.5 (DTXI), m/z 903.5 [right arrow] 137.5 (PTX2), and m/z 1141.5 [right arrow] 1061.5 (YTX), and calibration curves were constructed for all of the 4 parent compounds.
Yessotoxin (YTX), a disulfated polyether toxin was first isolated by Murata et al.
Standard solutions of OA (14.3 [micro]g/mL), PTX2 (8.6 [micro]g/mL), and YTX (5.3 [micro]g/mL) were obtained in methanol from the Institute for Marine Biosciences of National Research Council (Halifax, Nova Scotia).